1.0µg of E.coli in a standard sandwich ELISA. Assay by immunoelectrophoresis resulted in a single precipitin arc against purified and partially purified E.coli beta-galactosidase.

General notes

Application Note: This antibody shows inconsistent results between batches in ICC/IF, IF, IHC-P, IHC-Fr and IHC-FoFr. We have received several good Abreviews on these applications, however, we are unable to guarantee positive results. If the antibody is used for IHC, please try fixing with methanol and block with gelatine instead with BSA or NGS. After each antibody incubation, please wash 5x with TBS instead of PBS.We would recommend ab4761 or ab12081 as alternatives.

Properties

Form

Liquid

Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

Applications

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application

Abreviews

Notes

ELISA

1/10000.

sELISA tested against 1ug E.coli beta galactosidase.

WB

1/5000 - 1/10000. Detects a band of approximately 116 kDa.

This antibody shows inconsistent results between batches in ICC/IF, IHC-P, IHC-Fr and IHC-FoFr. We have received several good Abreviews on these applications, however, we are unable to guarantee positive results. If the antibody is used for IHC, please try fixing with methanol and block with gelatine instead with BSA or NGS. After each antibody incubation, please wash 5x with TBS instead of PBS. We would recommend ab4761 or ab12081 as alternatives.

IP

1/500 - 1/5000.

Application notes

Is unsuitable for ICC/IF,IHC-FoFr,IHC-Fr or IHC-P.

Target

Relevance

Beta galactosidase is coded by a gene (lac z) in the lac operon of Escherichia coli. It is a metalloenzyme that splits lactose into glucose and galactose. It hydrolyzes terminal, non-reducing beta-D-galactose residues in beta-D-galactosides.
Activation by cations seems to be substrate dependent. K+, Na+, NH4+, Rb+, Cs+ and Mn++ all activate enzyme activity based upon the substrate used.

Western blotting using ab616. Lane 1 shows 80 ng and lane 2 shows 20 ng loaded onto gel. Results for non-reducing conditions of SDS-PAGE prior to transfer to nitrocellulose are shown on the left side of the figure; results obtainined under reducing conditions are shown on the right. The membrane was probed with ab616 at 1/10,000 dilution.

I am very pleased to hear you would like to accept our offer and testab616 and ab15580in ICE. These codes will give you: 1 free primary antibody per code before the expiration date. To redeem this offer, please submit an Abreview for ICE for each antibody and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

Thank you for submitting an Abreview for ab616. As you may have noticed, your review has now been published on our website.Since you obtained poor results using the antibody in an untested species, we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.Based on the information in your review, I have a few more questions:1) Did you use the antibody to detect human beta Galactosidase ?2) If not, for which application or species was the discount code given?In terms of suggestions:- It might be helpful to compare the resuts obtained with human beta Galactosidase with E.coli beta Galactosidase. If you decide to follow these suggestions, please let me know if they are helpful.I wish you good luck with your research.

I am sorry to hear you are experiencing difficulties with one of our products. As discussed over the phone, I'd like to try and help you to improve the results currently obtained. In order to do this, could you please fill out the questionnair I have attached to this email. If you could also include some images of the blots obtained that would be very helpful. I understand that you have already performed a lotof optimisation but if you could just briefly outline what you have tried to optimise that would be very helpful.

I think it would be worthwhile to try and use the beta Galactosidase protein (ab7981) which you have just to check that there is not a problem with the antibody itself thatare contributing to the poor results.

In answer to you questions in regards to how to use this protein as a control. We recommend reconstituting the protein in a minimum concentration of 1 mg/mL in 0.1 M Sodium/potassium phosphate buffer, pH 7.0. This can be performed as follows:

The lypolised powder the protein is provided as can be stored at2-8°C but once dissolved it is best to aliquot and freeze stocks at -20C or -80C. We do not have any reconstituted stability data andwould therefore recommend to reconstitute the amount needed for the assay fresh just prior to performing the assay.

I would recommend initially loading 0.1 ug in a well to see how well the antibody performs next to your samples. This value may need to be reduced if the signal observed is too strong compared to your lysates.

I hope this information has been of help. I look forward to receiving your reply.